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Plasmids

Plasmids

Plasmids are extrachromosomal DNA molecules found in bacteria, archaea, and some eukaryotic microbes like yeast. These small, circular DNA structures typically contain fewer than 30 genes, although some may exist linearly. Plasmids vary in their number within a cell, known as copy number. Single-copy plasmids are present in one copy per cell and multi-copy plasmids are present in multiple copies, reaching over 100 copies per cell.Plasmids usually replicate independently of the chromosomal DNA...

DNA Bacteriophages

DNA Bacteriophages

Bacteriophages, or phages, are viruses that specifically infect bacteria, utilizing their genetic material to hijack host cellular machinery for replication. DNA bacteriophages employ single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA) genomes. These phages exhibit diverse replication strategies and host interactions, influencing their ecological roles and applications in biotechnology and medicine.ssDNA BacteriophagesssDNA phages, with their small genomes, utilize unique strategies to...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

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Updated: Sep 26, 2025

Quantification of Plasmid-Mediated Antibiotic Resistance in an Experimental Evolution Approach

Quantification of Plasmid-Mediated Antibiotic Resistance in an Experimental Evolution Approach

Published on: December 14, 2019

Evolution of plasmid-construction.

Xuegang Li¹, Jiacheng Jin¹, Zhanyong Guo²

¹The Life Science College, Henan Agricultural University, Zhengzhou 450002, China.

International Journal of Biological Macromolecules

|April 22, 2022

Summary

This summary is machine-generated.

Plasmid construction methods have evolved over 50 years, with 37 techniques now available. This review simplifies understanding and practice of these cloning strategies.

Keywords:

Annealing Evolution Mega-primer Plasmid-construction Recombination

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Last Updated: Sep 26, 2025

Quantification of Plasmid-Mediated Antibiotic Resistance in an Experimental Evolution Approach

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Area of Science:

Molecular Biology
Biotechnology
Genetic Engineering

Background:

Plasmid construction is fundamental to molecular biology and genetic engineering.
Over 37 distinct plasmid construction methods have been developed over the past 50 years.
Existing methods often present challenges in ease of use, efficiency, and seamless integration.

Purpose of the Study:

To provide a comprehensive global and evolutionary overview of plasmid construction methods.
To clarify and simplify the understanding and practical application of diverse cloning techniques.
To categorize existing methods based on underlying principles and mechanisms.

Main Methods:

Review and classification of 37+ plasmid construction methods.
Categorization based on three core principles: single-strand overhang, homologous recombination, and mega-primer amplification.
Analysis of evolutionary trends towards easier, more efficient, and seamless cloning.

Main Results:

Identified three primary cloning groups: single-strand overhang, homologous recombination, and mega-primer cloning.
Highlighted the evolutionary trajectory of methods aiming for improved cloning outcomes.
Recognized primer annealing and primer invasion as key underlying mechanisms.
Identified the scar junction as a common challenge in plasmid construction.

Conclusions:

A structured understanding of plasmid construction methods aids practical application.
Evolutionary trends indicate a continuous drive for enhanced cloning efficiency and simplicity.
Addressing challenges like scar junctions is crucial for advancing plasmid construction techniques.